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  • 1
    ISSN: 1432-0584
    Keywords: Elektronische Blutplättchenzählung ; Thrombozytopenien ; TOA-Thrombozytenzähler PL-100 ; plättchenreiches Plasma ; Vortex-Effekt ; Electronic platelet counting ; Thrombocytopenias ; TOA-platelet counter PL-100 ; Platelet-rich plasma ; Vortex-effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Platelet counts in platelet-rich plasma without hematocrit dependent correction were performed by following rapid and simple steps: l. pre-dilution of 20 μl of whole blood by an isotonic solution 1: 25; 2. stabilized low-speed centrifugation with 55 g for 5 minutes; 3. final dilution 1: 5000; 4. enumeration by use of a TOA platelet counter PL-100 which has been technically improved in comparison to similar machines. Erroneously high results were obtained after a too short or too low centrifugation. As reason for this artificial small pulses due to disturbances of the flow patterns around the aperture (so-called vortex-effect) can be assumed having been caused by large-volumed erythrocytes and leukocytes in the suspension. The routinely used procedure was reliable for all platelet ranges, especially in thrombocytopenias between 100 × 109/l and 25 × 109/l. In lower ranges comparisons with visual counts are essential.
    Notes: Zusammenfassung Thrombozytenzahlen wurden im plättchenreichen Plasma ohne Hämatokritkorrektur mit folgenden Schritten einfach und schnell ermittelt: 1. 1∶ 25-Vorverdünnung einer 20-μl-Vollblutprobe mit isotoner Lösung; 2. niedertourige stabilisierte Zentrifugation über 5 min bei 55 g; 3. Endverdünnung auf 1∶ 5000; 4. Zählung mit Hilfe eines gegenüber ähnlichen Geräten technisch verbesserten TOA-Thrombozytenzählers PL-100. Fälschlich erhöhte Zahlen wurden bei zu kurzer oder zu niedriger Zentrifugation gefunden. Ursächlich sind Störimpulse infolge Wirbelbildungen in der Umgebung der Meßkapillare anzunehmen (sog. Vortex-Effekt), die von verbliebenen größervolumigen Erythrozyten und Leukozyten in der Suspension hervorgerufen werden. Das routinemäßig angewandte Verfahren hat sich in allen Zählbereichen, insbesondere auch bei Thrombozytopenien zwischen 100 × 109/l und 25 × 109/l bewährt. In niedrigeren Bereichen sind Kontrollzählungen mit der Kammer erforderlich.
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  • 2
    ISSN: 1432-0584
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
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  • 3
    ISSN: 1432-0584
    Keywords: Cytogenetic studies ; AML ; Prognostic value
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Between 1981 and 1986 cytogenetic studies of bone marrow and/or blood cells in 139 patients with de novo acute myeloid leukemia (AML) were performed. The overall incidence of chromosomal aberrations was 53%, and this was not significantly influenced by sex, age nor the FAB classification. The aberrations most often found were: complex anomalies (n=13), t(8; 21) (n=10), trisomy 8 (n=9), monosomy 7 (n=6), monosomy 5, 5q-, trisomy 11, 12p- (n=4) and trisomy 6, 11q-, inv [16] (n=3). The prognostic significance of chromosomal findings was evaluated in 112 patients treated by combination chemotherapy. The chromosomal status NN, AN, AA did neither significantly influence complete remission rate (NN: 68%, AN: 71%, AA: 60%) nor mean survival (NN: 24, AN: 26.6, AA: 35.6 months). On the other hand, certain types of chromosomal anomalies were of prognostic value. CR was obtained in all 10 patients with t(8; 21) but only in 2 out of 9 patients with complex aberrations. Median duration of CR in patients with t(8; 21) was significantly longer than in patients with a normal karyotype (30 vs 7 months).
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  • 4
    ISSN: 1432-0584
    Keywords: Key words G-CSF ; AML ; Cytosine arabinoside ; Refractory disease ; Salvage therapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Patients with primary refractory or relapsed acute myeloid leukemia (AML) who undergo intensive salvage chemotherapy carry a high risk of treatment failure due to infectious complications and early relapses. The study presented here assessed the effect of granulocyte colony-stimulating factor (G-CSF) on the duration of post-treatment neutropenia, the incidence of infection-related deaths, and the disease-free and overall survival. Sixty-eight evaluable patients with relapsed and refractory AML received G-CSF 5 μg/kg per day subcutaneously starting 2 days after the completion of salvage treatment with the S-HAM regimen, consisting of high-dose cytosine arabinoside twice daily on days 1, 2, 8, and 9 and mitoxantrone on days 3, 4, 10, and 11. Ninety-one patients who were treated with the identical S-HAM regimen but without G-CSF support during a preceding study served as controls. The application of G-CSF resulted in a significant shortening of critical neutropenia of less than 500 μl (36 vs. 40 days;p=0.008), which translated into a trend towards a lower early death rate (21% vs. 30%) and an increase of complete remissions (56% vs. 47%, p=0.11). In patients younger than 60 years a significant prolongation of time to treatment failure (159 vs. 93 days, p=0.038) and of duration of disease-free survival (203 vs. 97 days, p=0.003) was observed. These results indicate a beneficial effect of G-CSF on early mortality as well as on long-term outcome when administered after S-HAM salvage therapy for advanced AML.
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  • 5
    ISSN: 1432-0584
    Keywords: Key words Cytogenetics ; Fibroblasts ; Hematopoietic microenvironment ; Leukemia ; Macrophages
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Bone marrow stroma cells from patients with acute myeloid leukemia (AML) display a variety of functional abnormalities. In order to determine whether this is related to an imbalance in the proportion of different stroma cell types or to integration of leukemic progeny into the regulatory cell network, stroma layers were established in mycophenolic acid-treated long-term marrow cultures from 16 patients with AML and 42 controls and analyzed by means of simultaneous membrane immunofluorescence and interphase cytogenetics. Macrophages were identified by CD14 expression, fibroblasts by staining with the AS02 antibody, and malignant cells by leukemia-specific numerical chromosome aberrations, including monosomy 7 and trisomy 8. Compared with normal controls, there was a slight decrease in the proportion of stroma fibroblasts (52±27% versus 77±5%) in 10-week-old cultures from patients with AML. Two of five AML patients with trisomy 8 and both patients with monosomy 7 had evidence of leukemic stroma cells. Most malignant cells were CD14+ macrophages (3.8–98.1% of all CD14+ cells), but some were AS02+ (2.8–5.2%). AML stroma layers showed a reduced capacity to support the growth of normal hematopoietic cells in standard two-stage long-term cultures, but this was unrelated to the presence or absence of leukemic stroma elements. In conclusion, AML populations vary with respect to their ability to produce a malignant microenvironment. Functional defects in the hematopoietic microenvironment, however, are not limited to AML patients with cytogenetically abnormal stroma cells, but extend to cases without evidence of malignant stroma cells.
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  • 6
    ISSN: 1432-0584
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
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  • 7
    ISSN: 1432-0584
    Keywords: Lymphocyte volume ; Electronic modal volume ; Chronic Lymphocytic leukemia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The electronic modal lymphocyte volumes of 151 patients with chronic lymphocytic leukemia (CLL) and 305 normal controls were determined by the hydrodynamically focused multi-channel Coulter TF analyser. The mean volumes of the normally distributed groups were 166±19.3 (range 126–216) fl in patients with CLL and 206±14.4 (range 126±246) fl in normal controls. The calculated cell diameters were 6.8 (6.2–7.4) μm and 7.3 (6.8–7.8) μm respectively. Our data do not support previous reports about relations between cell size and clinical stages of the Rai and Binet classifications.
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  • 8
    ISSN: 1432-0584
    Keywords: Silver staining ; Ferritin ; Transmission electron microscopy ; X-ray microanalysis ; Iron overload
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A short exposure of cell suspensions to gaseous hydrogen sulfide, appropriate fixations, and subsequent physical development of silver shells around sulfidated insoluble metals were used to amplify ferritin iron cores in blood and bone marrow cells. The methods described are suitable for both light microscopy and transmission electron microscopy. These techniques made it possible to visualize Prussian Blue stainable ferritin and haemosiderin, as well as a large variety of isoferritin iron and other smaller particles beyond the sensitivity of Prussian Blue staining. Admixtures of sulfidatible zinc and traces of other heavy metals had to be taken into consideration. For further research, adaptations of sulfide silver staining to microphysical analyses were developed. However, conventional energy dispersive X-ray analysis was not sensitive enough to signalize the presence of Fe in sulfide silver amplified iron cores of a single or a few ferritin molecule(s). Proton-induced X-ray emission was used to measure Fe and Zn down to 1 fg/single cell in unstained or sulfide silver stained smears on thin foils. However, multielement analysis of homogeneous cell concentrates was much easier to perform and far more sensitive. In advanced iron overload, highly increased sulfide silver staining was found in peripheral blood cells including lymphocytes, monocytes, eosinophils, basophils, and — in extreme cases — also in neutrophils and platelets.
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  • 9
    ISSN: 1432-0584
    Keywords: Ph1+-ALL ; Prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Between 1983 and 1991 the Philadelphia chromosome (Ph1) was found in bone marrow and/or peripheral blood cells of 25 adult patients with acute lymphoblastic leukemia (ALL). The Ph1 as sole anomaly was seen in 13 patients, while six patients had additional structural and another six structural and numerical aberrations. Most patients (23/25) received combination chemotherapy according to the BMFT protocols 1/81, 2/84, 3/87, and 4/89. For 25 evaluable patients two early deaths, two treatment failures, two partial remissions (PR), and 19 complete remissions (CR) after phase 1 or 2 of the induction regimen were recorded. Two of these 19 patients who achieved CR are presently disease free, whereas 17 have relapsed after a median duration of remission of 9 months. Actuarial median survival for all patients was 13 months. The probability of continuous complete remission (CCR) after 39 months, as well as that of survival after 40 months, is only 6%. Our results confirm that the presence of the Ph1 is associated with a poor prognosis in adult-ALL patients. Therefore, whenever first CR is obtained and an HLA-identical donor is available, allogeneic bone marrow transplantation (BMT) should be performed at once, the more so, since transplantation in second CR seems to offer no cure. Future studies will have to show whether an intensified cytotoxic therapy can improve the prognosis of Ph1+-ALL.
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  • 10
    ISSN: 1432-0584
    Keywords: Reticulocytes ; Automatic counters ; Acute myeloid leukemia ; Bone marrow recovery
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Flow-cytometric reticulocyte counts including their maturation status were performed during follow-up of induction (n=5) and consolidation (n=7) polychemotherapy in nine patients with acute myeloid leukemia (AML) using a Sysmex R-3000 automated counter. The reticulocytes fell to an extreme nadir (〈0.01/pl) — as did leukocytes and platelets — and consisted of cells with low fluorescence ratio (LFR) only. After a median interval of 16 days, the fraction with medium fluoresence ratio (MFR) began to rise, preceding the reticulocytes with high fluorescence ratio (HFR) for a median of 1 day in all cases with partial (n=1) or complete (n=8) remission. At a median of 7 days after MFR and 5 days after HFR the granulocytes reached the critical limit of ≥0.5/nl. The reticulocytes rose to the normal range after 9 and 7 days, respectively. Automated flow-cytometric reticulocyte counting including the maturation status has been shown to provide an accessible measure of erythroid activity, which may be of predictive value for granulocyte recovery after aplasiogenic polychemotherapy in AML patients.
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